Skin tape stripping: a non-invasive diagnostic strategy for dermal exposure to cytotoxic compounds

Abstract

A non-invasive method for determining a subject's dermal exposure to an agent employing an immunohistochemical procedure on a skin strip applied to an exposed area on the subject's person and detecting agent adduct thereon.

1 . A polyclonal antibody to sulfur mustard adducts purified from mammalian antisera, said antisera derived from the protocol described in Table B herein using mammals immunized against said sulfur mustard adducts. 
   
   
       2 . The polyclonal antibody of  claim 1 , wherein said sulfur mustard adducts are derived from keratinocytes whole cell lysates. 
   
   
       3 . A non-invasive method for determining a subject's dermal exposure to an agent, wherein said method comprises:
 applying a transparent adhesive tape to a target area of the skin of the subject in a manner sufficient to isolate an epidermal sample adhering to the adhesive tape, wherein the epidermal sample comprises cells from the stratum corneum of the subject;   employing an immunohistochemical procedure on the sample present on said tape resulting in a treated sample, said immunoshitochemical procedure employing an antibody specific to the adducts of said agent; and   examining the treated sample present on said tape to determine the presence of said agent adduct therein.   
   
   
       4 . The method of  claim 3 , said transparent tape is double sided, with adhesive on both sides of said transparent double sided tape. 
   
   
       5 . The method of  claim 3 , wherein said immunohistochemical procedure is indirect immunoperoxidase procedure. 
   
   
       6 . The method of  claim 3 , wherein said antibody is a polyclonal antibody. 
   
   
       7 . The method of  claim 3 , wherein said antibody is a monoclonal antibody. 
   
   
       8 . The method of  claim 3 , wherein said antibody is a recombinant antibody. 
   
   
       9 . The method of  claim 3 , wherein said antibody is a phage antibody. 
   
   
       10 . The method of  claim 3 , wherein said antibody is a synthetic protein constructed on a non-immunoglobulin scaffold selected from the group consisting of protein A, lipocalins, fibronectin domains, ankryin consensus repeat domains and thioredoxin. 
   
   
       11 . The method of  claim 6 , wherein the polyclonal antibody is purified from mammalian antisera obtained in the protocol described in Table B herein. 
   
   
       12 . The method of  claim 4 , wherein the double sided transparent tape is selected from the group consisting of tapes described in Table B herein. 
   
   
       13 . The method of  claim 4 , wherein the double sided transparent tape comprises a soft and pliable polyethylene backing layered with an acrylate based adhesive. 
   
   
       14 . The method of  claim 3 , wherein said antibody is specific to keratin adducts formed following mustard sulfur exposure. 
   
   
       15 . The method of  claim 3 , wherein said treated sample is examined using a device selected from the group consisting of a densitometer, a reflectometer, a light microscope and a calorimeter. 
   
   
       16 . The method of  claim 3 , wherein said agent is selected from the group consisting of sulfur mustard, nitrogen mustard, and lewisite. 
   
   
       17 . A kit for use in determining whether a subject has been exposed to an agent, wherein said kit comprises
 at least one positively charged microscope slide;   mounting media;   glass cover slips;   nonspecific staining solutions;   double-sided transparent tape suitable to obtain a skin sample from said subject;   indirect immunoperoxidase lyophilized primary antibody specific to adducts formed by said agent;   a secondary antibody;   avidin-biotin complex;   diaminobenzidine chromogen tablets;   case designed to accommodate all said kit components.   
   
   
       18 . The kit of  claim 17 , wherein said primary antibody is specific to keratin adducts formed following sulfur mustard exposure. 
   
   
       19 . The kit of  claim 17 , wherein said primary antibody is a polyclonal antibody purified from mammalian antisera, said antisera derived from the protocol described in Table B herein using mammals immunized against sulfur mustard adducts. 
   
   
       20 . The kit of  claim 17 , wherein said kit further comprises a device selected from the group consisting of a densitometer, a reflectometer, a light microscope and a calorimeter. 
   
   
       21 . The kit of  claim 17 , wherein said kit further comprises instructions for use of said kit in determining whether said subject has been exposed to said agent. 
   
   
       22 . The kit of  claim 17 , wherein the agent is selected from a group consisting of sulfur mustard, nitrogen mustard, and lewisite.